CA2446157A1 - Methods for screening substances in a microwell array - Google Patents

Methods for screening substances in a microwell array Download PDF

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Publication number
CA2446157A1
CA2446157A1 CA 2446157 CA2446157A CA2446157A1 CA 2446157 A1 CA2446157 A1 CA 2446157A1 CA 2446157 CA2446157 CA 2446157 CA 2446157 A CA2446157 A CA 2446157A CA 2446157 A1 CA2446157 A1 CA 2446157A1
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Prior art keywords
liquid
holes
platen
containers
distinct
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CA 2446157
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French (fr)
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CA2446157C (en
Inventor
Ian W. Hunter
Colin J. H. Brenan
Tanya S. Kanigan
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Massachusetts Institute of Technology
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Individual
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/02—Drop detachment mechanisms of single droplets from nozzles or pins
    • B01L2400/022—Drop detachment mechanisms of single droplets from nozzles or pins droplet contacts the surface of the receptacle
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L9/00—Supporting devices; Holding devices
    • B01L9/52—Supports specially adapted for flat sample carriers, e.g. for plates, slides, chips
    • B01L9/523—Supports specially adapted for flat sample carriers, e.g. for plates, slides, chips for multisample carriers, e.g. used for microtitration plates
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B60/00—Apparatus specially adapted for use in combinatorial chemistry or with libraries
    • C40B60/14—Apparatus specially adapted for use in combinatorial chemistry or with libraries for creating libraries
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B70/00—Tags or labels specially adapted for combinatorial chemistry or libraries, e.g. fluorescent tags or bar codes
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N2035/00178—Special arrangements of analysers
    • G01N2035/00237—Handling microquantities of analyte, e.g. microvalves, capillary networks
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N2035/1027—General features of the devices
    • G01N2035/1034—Transferring microquantities of liquid
    • G01N2035/1037—Using surface tension, e.g. pins or wires
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/26—Conditioning of the fluid carrier; Flow patterns
    • G01N30/38—Flow patterns
    • G01N30/46—Flow patterns using more than one column
    • G01N30/466—Flow patterns using more than one column with separation columns in parallel
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/60—Construction of the column
    • G01N30/6095—Micromachined or nanomachined, e.g. micro- or nanosize
    • G—PHYSICS
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    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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    • G01N35/00029—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor provided with flat sample substrates, e.g. slides
    • G01N35/00069—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor provided with flat sample substrates, e.g. slides whereby the sample substrate is of the bio-disk type, i.e. having the format of an optical disk
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
    • Y10T436/00—Chemistry: analytical and immunological testing
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    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
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    • Y10T436/00—Chemistry: analytical and immunological testing
    • Y10T436/25—Chemistry: analytical and immunological testing including sample preparation
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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    • Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
    • Y10T436/00—Chemistry: analytical and immunological testing
    • Y10T436/25—Chemistry: analytical and immunological testing including sample preparation
    • Y10T436/25625—Dilution
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
    • Y10T436/00—Chemistry: analytical and immunological testing
    • Y10T436/25—Chemistry: analytical and immunological testing including sample preparation
    • Y10T436/2575—Volumetric liquid transfer
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
    • Y10T83/00—Cutting
    • Y10T83/04—Processes
    • Y10T83/0481—Puncturing

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  • Chemical & Material Sciences (AREA)
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  • Biochemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Physics & Mathematics (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Automatic Analysis And Handling Materials Therefor (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Feeding, Discharge, Calcimining, Fusing, And Gas-Generation Devices (AREA)
  • Physical Or Chemical Processes And Apparatus (AREA)
  • Micromachines (AREA)
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  • Sampling And Sample Adjustment (AREA)

Abstract

Methods for manufacturing and using an apparatus for manipulating and analyzing a large number of microscopic samples of a liquid or materials, including cells, in liquid suspension. Parallel through-holes (12) are formed in a platen (10) and loaded with a liquid. Loading may be performed in such a way as to create a gradient, with respect to the position of the through-holes, of the concentration of a particular substance or of another quantity.
Mixing of the contents of through-holes may be obtained by bringing filled microwell arrays into contact with each other with registration of individuals through-holes.

Claims (26)

1. A method for loading a plurality of disparate sample containers, the sample containers comprising an integral structure, such that the concentration of a specified substance in each container is characterized by a gradient with respect to position of the container within the structure, the method comprising:
a. introducing a first liquid into the disparate containers; and b. contacting the containers with a second liquid, the second liquid containing the specified substance, in such a manner that the degree of diffusion of the specified substance into the disparate containers differs in a controlled manner among the containers.
2. A method according to claim 1, wherein contacting the containers with the second liquid includes differing a duration of contact with the second liquid as among the disparate containers.
3. A method according to claim 1, wherein contacting the containers with the second liquid includes modulating the diffusion rate of the specified substance into the containers.
4. A method according to claim 3, further comprising the step of creating a gradient of the specified substance in the second liquid prior to contacting the containers with the second liquid.
5. A method according to claim 4, wherein the step of creating a gradient of the specified substance includes applying electrophoresis.
6. A method according to claim 3, wherein the step of modulating the diffusion rate includes contacting the containers with the second liquid through a membrane having a permeability variable with respect to position.
7. A method according to claim 1, wherein the step of introducing a first liquid into the disparate containers includes filling a platen having a plurality of through-holes.
8. A method for loading a liquid into a plurality of through-holes of a platen, the platen having a top surface, the top surface having an opening to each through-hole, the method comprising:
a. forming a droplet of the liquid on the end of a liquid transfer device;
b. moving the fluid transfer device so as to drag the droplet across the top surface of the platen and over the through-holes to be filled;
c. dispensing the liquid in such a manner as to keep the droplet from being depleted;
and d. withdrawing the droplet from the surface after the through-holes are filled.
9. A method for loading a liquid sample into a plurality of through-holes of a platen, the method comprising:
a. filling a plurality of capillary tubes having dispensing ends, the capillary tubes comprising an array;
b. disposing each dispensing end in proximity to a distinct through-hole; and c. transferring liquids to the through-holes of the platen through the capillary tubes.
10. A method for loading a plurality of sample containers, the sample containers comprising an integral structure, such that the concentration of a specified substance in each container is characterized by a gradient with respect to position of the container within the structure, the method comprising:
a. creating regions of distinct concentration of the specified substance in a liquid, the liquid having a surface; and b. transferring into distinct containers the liquid from the distinct regions.
11. A method according to claim 10, wherein the step of creating regions of distinct concentration includes applying electrophoresis.
12. A method according to claim 10, wherein the step of transferring liquid includes transporting the liquid through capillary tubes.
13. A method according to claim 10, wherein the step of transferring the liquid includes contacting the integral structure with the surface of the liquid.
14. A method for manufacturing a platen having two substantially parallel planar surfaces and a plurality of through-holes disposed substantially perpendicularly to the planar surfaces, the method comprising:
a. providing a sheet of thermoplastic material;
b. loading the sheet of thermoplastic material into contact with a surface of a die having a plurality of holes; and c. bringing a punch having a plurality of protrusions of specified cross-section into contact with the sheet of thermoplastic material in such a manner that the protrusions are in alignment with the holes of the die such that through-holes are cut through the thermoplastic material.
15. A method for manufacturing a platen having two substantially parallel planar surfaces and a plurality of through-holes disposed substantially perpendicularly to the planar surfaces, the method comprising:
a. providing a sheet of electrically conducting material;
b. loading the sheet of conducting material into contact with a surface of a die having a plurality of holes; and c. bringing an EDM mandrel having a plurality of protrusions of specified cross-section into proximity of the sheet of conducting material in such a manner that the protrusions are in alignment with the holes of the die such that through-holes are cut through the conducting material.
16. A method for providing a hydrophobic coating to a silicon platen having a first and a second surface, the surfaces being substantially parallel, and a plurality of through-holes substantially perpendicular to the surfaces, the method comprising:
a. oxidizing the first surface;
b. cleaning the oxidized first surface;
c. applying a positive pressure of inert gas to the plurality of through-holes from the direction of the second surface; and d. exposing the first surface to a silanizing vapor agent.
17. A method for loading a liquid into a plurality of through-hole platens, each platen having a plurality of through-holes, the method comprising:
a. stacking at least two platens together in such an adjacent manner that at least one of the plurality of through-holes from each platen is registered with a through-hole of each other adjacent platen so as to form at least one continuous channel;
and b. transferring the liquid into each continuous channel.
18. A method according to claim 17, wherein each platen is separated from each adjacent platen by an air gap.
19. A method according to claim 17, wherein the step of transferring liquid includes transporting the liquid through capillary tubes.
20. A method according to claim 17, wherein the step of transferring liquid includes transporting the liquid through at least one cannula.
21. A method for mixing liquid in at least two platens, each platen having a plurality of through-holes, the method comprising stacking at least two platens together for a specified time, in such a manner that at least one of the plurality of through-holes from each platen connects with a corresponding through-hole of another platen such that liquid is allowed to diffuse between connecting through-holes.
22. A method according to claim 21, further including the step of separating the at least two platens after mixing of the liquid between the connecting through-holes.
23. A microlens array for diffusing light, the microlens array comprising, a. a microchannel plate having a plurality of parallel microchannels; and b. a liquid disposed within substantially each of the plurality of the parallel microchannel.
24. A method for humidifying a system, the method comprising, a. filling a microchannel plate with a liquid having a plurality of parallel microchannels; and b. placing the filled microchannel plate in vicinity of the system to be humidified.
25. A method for diffusing light, the method comprising:
a. entraining a fluid in substantially each of a plurality of parallel microchannels having proximal and distal ends, the microchannels comprising a microchannel plate;
b. illuminating the proximal end of each of the plurality of parallel microchannels with light; and c. allowing diffuse light to emanate from the distal ends of the microchannel.
26. A perforated platen having substantially parallel planar surfaces for manipulating distinct liquid samples, each sample having a volume less than 1 microliter, the platen comprising:
a. an inner layer of hydrophilic material;
b. two outer layers of hydrophobic material coupled to opposite sides of the inner layer; and c. a two-dimensional array of through-holes, at least two holes having distinct volumes, for retaining the distinct liquid samples, the through-holes each having a diameter less than 300 micrometers and traversing the inner layer and the two outer layers in a direction substantially perpendicular to the planar surfaces of the platen.
CA 2446157 2001-05-07 2002-05-07 Methods for screening substances in a microwell array Expired - Lifetime CA2446157C (en)

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US09/850,123 US6893877B2 (en) 1998-01-12 2001-05-07 Methods for screening substances in a microwell array
US09/850,123 2001-05-07
PCT/US2002/014357 WO2002089982A2 (en) 2001-05-07 2002-05-07 Methods for screening substances in a microwell array

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